Brains and spinal cords were dissected and post-fixed overnight in paraformaldehyde, then transferred to 30% sucrose/PBS solution for 2 d. 50-µm-thick coronal sections of the cervical enlargement were cut using a cryostat, collected, and processed free-floating. To visualize CST collaterals, the BDA signal was amplified by incubation with ABC Complex (Vector Laboratories) overnight at 4°C. After a 30-min tyramide amplification (Biotin-XX; TSA Kit 21; Life Technologies), sections were incubated overnight with streptavidin-conjugated Alexa Fluor 647 or streptavidin-conjugated Alexa Fluor 594 (1:500; Life Technologies).
Biotin xx tsa kit 21
Manufactured by Thermo Fisher Scientific
Biotin-XX; TSA Kit 21 is a laboratory product designed for use in biotech and life science research applications. It serves as a detection system for target biomolecules. The kit includes necessary reagents and components to facilitate the detection process.
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2 protocols using biotin xx tsa kit 21
1
Visualizing Corticospinal Tract Collaterals
2
Visualizing Corticospinal Tract Collaterals
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Brains and the cervical enlargement C3‐C5 of the spinal cords were dissected and postfixed overnight in PFA, then transferred to 30% sucrose/PBS solution overnight. 30 μm thick coronal sections of the cervical enlargement was cut using a cryostat, collected, and processed free floating. To visualize CST collaterals, the BDA signal was amplified by incubation with ABC Complex (Vector Laboratories) overnight at 4°C. After a 30‐min tyramide amplification (Biotin‐XX, TSA Kit #21, Life Technologies), sections were incubated overnight with Streptavidin‐conjugated Alexa Fluor 647 or Streptavidin‐conjugated Alexa Fluor 594 (1:500, Life technologies).
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